Objective To explore the effect of multi-sector cooperation management mode on standardizing submission of specimens for blood culture and application of antimicrobial agents. Methods Blood culture specimens from hospitalized patients in a tertiary first-class hospital from 2017 to 2018 were selected as research data. January-December 2017 was as pre-intervention group and January-December 2018 was as post-intervention group. From December 2017, a multi-sector cooperation management team was established, and other intervention measures were gradually implemented. Submission rate, submission mode, contamination rate, and positive rate of blood culture specimens, as well as antimicrobial use rate in hospitalized patients before and after the implementation of intervention of multi-sector cooperation management mode were compared. Results After intervention, submission rate of blood specimens of the whole hospital before therapeutic, restrictive and special-class antimicrobial use were 22.29%, 30.16% and 60.30% respectively, which were higher than 14.91%, 18.55% and 47.05% before intervention, difference were all significant (all P<0.001). The proportion of double and above sets blood culture increased from 15.48% before intervention to 26.97% after intervention. Contamination rate of blood culture after intervention was lower than before intervention (1.02% vs 2.98%), and positive rate of blood culture was higher than before intervention(11.67% vs 9.52%). Antimicrobial use rate after intervention was lower than that before intervention (34.11% vs 37.88%); use rates of restrictive antimicrobial agents and special-class antimicrobial agents after intervention were 14.67% and 2.56% respectively, which were lower than 15.60% and 3.73% respectively before intervention, differences were both significant(both P<0.001). There was significant difference in the constituent of combined use of therapeutic antimicrobial agents (P<0.001). Conclusion Through multi-sector cooperation management mode combining quality control and administrative management, submission quality of blood culture is effectively improved.